Please use this identifier to cite or link to this item: http://rportal.lib.ntnu.edu.tw:80/handle/20.500.12235/111524
Title: 探討海洋深層水對痤瘡丙酸桿菌誘發ICR小鼠耳朵皮膚發炎之改善效果
Effects of deep sea water on Cutibacterium acnes-induced skin inflammation in ICR mice ears
Authors: 鄭劍廷
Chien, Chiang-Ting
翁于晴
Weng, Yu-Ching
Keywords: 痤瘡丙酸桿菌
海洋深層水
發炎
ICR小鼠
p38
NF-kB
Cutibacterium acnes
deep sea water
inflammation
ICR mice
p38
NF-kB
Issue Date: 2019
Abstract: 痤瘡丙酸桿菌 (Cutibacterium acnes, P. acnes) 是一種兼性厭氧的革蘭氏陽性菌,常見於人體皮膚的皮脂腺上,然而當痤瘡丙酸桿菌不正常增生的情況下與致痤瘡的形成有關,是因為痤瘡丙酸桿菌在痤瘡的形成中,扮演促進發炎的重要角色。海洋深層水 (Deep sea water, DSW) 含有豐富礦物質與活性成分,有文獻指出海洋深層水可以預防或治療過敏性濕疹,然而目前對於海洋深層水治療痤瘡丙酸桿菌所引起的痤瘡效果未有深入的研究。因此本研究探討海洋深層水對痤瘡丙酸桿菌誘導小鼠耳朵發炎情形是否有改善進行探討。首先在體外試驗中以紙錠擴散法 (disk diffusion method) 測定海洋深層水對痤瘡丙酸桿菌的抗菌活性,加入海洋深層水的載體與加入二次水的載體凝膠進行對照,可得知海洋深層水凝膠的紙錠周圍所生長的菌落數比二次水凝膠的紙錠周圍菌落數較少,因此在體外試驗具有抑菌的效果。動物實驗部分以ICR小鼠做為實驗對象,小鼠在經過一週飼養後,將痤瘡丙酸桿菌 6*109 CFU/uL 注射於小鼠耳朵皮下組織,誘導組織形成發炎現象,接著連續5天給予塗抹海洋深層水凝膠於耳朵誘發處,透過雷射都普勒血流儀觀察,予海洋深層水凝膠的組別耳朵降低紅腫的情況明顯好於其他組,且耳朵的腫脹程度也有明顯的下降。此外在H&E染色下,可以觀察到經過海洋深層水治療後可以減少皮下組織的發炎細胞浸潤的現象。我們再以西方墨點法觀察痤瘡丙酸桿菌對組織造成發炎反應之路徑,其主要是藉由活化p38及NF-kB磷酸化蛋白,進而產生發炎現象,但在給予海洋深層水後可抑制磷酸化蛋白的表現。另外在海洋深層水抑制促發炎因子的部分,包括granulocyte- macrophage colony-stimulating factor (GM-CSF)、Chemokine (C-C motif) ligand 5 (CCL5)、Chemokine (C-X-C motif) ligand 1(CXCL1)、interleukin-1b (IL-1b)、Chemokine (C-X-C motif) ligand 11 (CXCL11) 等促發炎因子之蛋白表現量有減少的趨勢。總結來說我們的結果表示經由海洋深層水治療後,能抑制MAPK 中的p38和NF-kB蛋白的磷酸化,並減少促發炎因子GM-CSF、 CXCL1、CCL5、CXCL11、IL-1b的產生,來達到降低P. acnes誘導組織發炎反應的情形,改善痤瘡丙酸桿菌引起的皮膚炎症之效果。因此海洋深層水能有效治療P. acnes誘導的發炎情形,以後可以作為痤瘡桿菌的治療選擇之一。
Cutibacterium acnes (Propionibacterium acnes, P. acnes) is a Gram-positive anaerobic bacterium exist in sebaceous follicle-rich areas of human skin. The overgrowth of P. acnes has been associated with the progression of acne. It plays an important role in the pathogenesis of acne by inducing certain inflammatory mediators. Deep sea water (DSW) is rich in minerals and active ingredients. DSW has displayed an effective potential in anti-atopic dermatitis activity in previous studies. Until now, the anti-P. acnes. activity against has not been clearly reported. This study investigated the effect and mechanism of DSW on anti-P. acnes induced inflammation. In this study, the DSW gel showed a good anti-microbial activity than the water gel in disk diffusion method. It means DSW gel had less microbiological colony count than water gel. Moreover the ICR mice were tested in this experiment. After 1 week of adaptation, we injected P. acnes (6*109 CFU/uL) into the ears induce inflammatory response. We applied the ears with DSW gel for 5 days after inflammation, followed by sacrifice and collected the tissue for subsequent experimental analysis. Our data showed that P. acnes enhanced blood flow in the mice ears. Treatment of DSW gel reduced inflammatory ears blood flow. In H&E stain, DSW gel also reduced ear inflammatory cell infiltration induced inflammation. We used western blot to further determine which pathway responsible for P. acnes induced inflammation. The data showed that P. acnes increased the expressions of phosphorylated nuclear factor-kB (NF-kB) and p38 in P. acnes- treated ears. DSW gel- treated ears displayed a downregulation of phosphorylated nuclear factor-B (NF-B) and p38 protein. DSW also exhibited an anti-inflammatory role by inhibiting the expression of pro-inflammatory cytokines, including granulocyte- macrophage colony-stimulating factor (GM-CSF), Chemokine (C-C motif) ligand 5 (CCL5), Chemokine (C-X-C motif) ligand 1 (CXCL1), interleukin-1b (IL-1b), Chemokine (C-X-C motif) ligand 11 (CXCL11). In summary, our results showed that DSW gel could suppress the inflammation through inhibition of p38/NF-kB pathway and reduced pro-inflammatory cytokines including GM-CSF, CXCL1, CCL5, CXCL11 and IL-1b expression. DSW gel showed a good effect on anti-P. acnes induced inflammatuib in the ears. DSW gel can provide a therapeutic potential on treating dermatitis.
URI: http://etds.lib.ntnu.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=id=%22G060643040S%22.&%22.id.&
http://rportal.lib.ntnu.edu.tw:80/handle/20.500.12235/111524
Other Identifiers: G060643040S
Appears in Collections:學位論文

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